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Biomedical subjects

M F Zhang

Publications and source records attributed to M F Zhang.

At least 19 recordsLinked to original sources
PubMed2006

Transcriptional profiles of chicken embryo cell cultures following infection with infectious bursal disease virus.

Infectious bursal disease virus (IBDV) is the causative agent of infectious bursal disease in chickens and causes a significant economic loss for the poultry industry. Little is understood about the mechanism involved in the host responses to IBDV infection. For better understanding the IBDV-host interaction, we measured steady-state levels of transcripts from 28 cellular genes of chicken embryo (CE) cell cultures infected with IBDV vaccine stain Bursine-2 during a 7-day infection course by use of the quantitative real-time RT-PCR SYBR green method. Of the genes tested, 21 genes (IRF-1, IFN 1-2 promoter, IFNAR-1, IRF-10, IFN-gamma, 2',5'-OAS, IAP-1, caspase 8, TRAIL-like, STAT-3, IL-6, IL-8, MIP-3 alpha, MHC-I, MHC-II, TVB, GLVR-1, OTF, IL-13R alpha, ST3GAL-VI and PGK) showed an increased expression. The remaining seven genes (IFNAR-2, IFN-alpha, NF-kappaB subunit p65, BLRcp38, DDX1, G6PDH and UB) showed a constant expression or only slight alteration. Apparently, the host genes involved in pro-inflammatory response and apoptosis, interferon-regulated proteins, and the cellular immune response were affected by IBDV infection, indicating involvement in the complex signaling pathways of host responses to the infection. This study thus contributes to the understanding of the pathogenesis of IBD and provides an insight into the virus-host interaction.

Animals↗
PubMed2006

Relative quantification and detection of different types of infectious bursal disease virus in bursa of Fabricius and cloacal swabs using real time RT-PCR SYBR green technology.

In present study, different types of infectious bursal disease virus (IBDV), virulent strain DK01, classic strain F52/70 and vaccine strain D78 were quantified and detected in infected bursa of Fabricius (BF) and cloacal swabs using quantitative real time RT-PCR with SYBR green dye. For selection of a suitable internal control gene, real time PCR parameters were evaluated for three candidate genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 28S rRNA and beta-actin to IBDVs. Based on this beta-actin was selected as an internal control for quantification of IBDVs in BF. All BF samples with D78, DK01 or F52/70 inoculation were detected as virus positive at day 1 post inoculation (p.i.). The D78 viral load peaked at day 4 and day 8 p.i., while the DK01 and F52/70 viral load showed relatively high levels at day 2 p.i. In cloacal swabs, viruses detectable were at day 2 p.i. for DK01 and F52/70, day 8 p.i. for D78. Importantly, the primers set were specific as the D78 primer set gave no amplification of F52/70 and DK01 and the DK01 primer set gave no amplification of D78, thus DK01 and D78 could be quantified simultaneously in dually infected chickens by use of these two set of primers. The method described here is robust and may sever as a useful tool with high capacity for diagnostics as well as in viral pathogenesis studies.

Actins↗
PubMed2005

Comparison of the survival of human biopsied embryos after cryopreservation with four different methods using non-transferable embryos.

BACKGROUND: The standard embryo cryopreservation method is still less than optimal for biopsied embryos. The aim of this study was to compare the survival of biopsied embryos cryopreserved with four different methods using non-transferable embryos. METHODS: Abnormal embryos from one or three pronuclei and spare embryos of grade 3 and 4 were used for this study. Non-biopsied embryos were cryopreserved using the standard method as control. Biopsied embryos were cryopreserved using four methods as follows: standard method, modified freezing method, modified thawing method and vitrification. Blastomere survival and blastulation of frozen-thawed embryos were compared between the different methods. RESULTS: The proportion of embryos with > or = 50% blastomere survival and total blastomere survival rate of biopsied embryos were significantly higher with vitrification than the other three methods. Both the modified freezing and modified thawing methods had significantly higher embryo survival and total blastomere survival rates than standard methods. However, there was no significant difference in blastulation of surviving embryos in all the five groups. CONCLUSIONS: Non-transferable embryos derived from clinical IVF/ICSI are useful for evaluation of the optimal freezing procedures for biopsied embryos. Vitrification increases the survival rate of human biopsied embryos above standard and modified cryopreservation methods.

Biopsy↗
PubMed2004

Rapid pathotyping of Newcastle disease virus from allantoic fluid and organs of experimentally infected chickens using two novel probes.

A system including RNA isolation, primers and two novel oligonucleotide probes (NC22 and VC22) was established to rapidly pathotype Newcastle disease virus (NDV) from infected allantoic fluid; The sequence of the probes is: VC22, 5'-AAGGAGGCAGAAACGCTTTATA-3'; NC22, 5'-GGGGAGACAGG GGCGCCTTATA-3'. Application efficacy of the probes was verified by differentiating NDV that derived from experimentally infected organs. NC22 and VC22 both were labeled with digoxigenin (DIG) and were successful in differentiating NDV strains from the infected allantoic fluid and organs of experimentally infected SPF chickens. The RT-PCR products of NDV isolates and strains were dotted onto nylon membrane and then hybridized with two specific probes separately. The VC22 probe is specific to virulent strains, and the NC22 probe is specific to avirulent strains. The results of hybridization coincide with viral intracerebral pathologenicity index (ICPI). The specificity of two probes and sensitivity of NC22 probe were evaluated. NC22 could detect down to 10(-8) dilution from 10(7.5) EID(50)/ml or 800 fg of viral RNA. The system could be completed within 1 day to conduct experiment from clinical sample to the result of assay, and its potential practical application in clinical assay was discussed basing on specificity, sensitivity and rapidness.

Allantois↗
PubMed2004

Assessment of genetic, antigenic and pathotypic criteria for the characterization of IBDV strains.

The aim of this work was the selection and comparison of representative infectious bursal disease virus (IBDV) strains. Nine strains of IBDV, isolated at different times and from different geographic regions of Europe and China, were characterized. Batches of all strains were prepared following standardized protocols and checked for the absence of contaminating viruses. Criteria used for their characterization were: (i) the nucleotide sequence of the VP2 variable region, (ii) binding to a panel of neutralizing monoclonal antibodies in antigen capture enzyme-linked immunosorbent assays, and (iii) virulence in specific pathogen free chickens after infection with a standardized number of median embryo infective doses. Based on the first two criteria, two of nine strains were classified as classical virulent (cv) IBDV (F52/70, Cu-1wt), and five as very virulent (vv) IBDV (849VB, 96108, HK46, GX, Harbin). Remarkably, although a clear-cut difference was demonstrable between European cvIBDV (F52/70 and Cu-1wt) and vvIBDV (849VB and 96108) strains, there was a continuum in the pathogenicity of Chinese vvIBDVs. Our results indicate the probable existence of differences in virulence within IBDV lineages determined on the basis of antigenic typing using monoclonal antibodies and the alignment of the VP2 sequences. This indicates limitations in the analysis of IBDV pathotypes based on the VP2 variable region and emphasizes that these criteria may not be sufficient for the classification of IBDV strains.

Amino Acid Sequence↗
PubMed2002

Early stages of infectious bursal disease virus infection in chickens detected by in situ reverse transcriptase-polymerase chain reaction.

Two infectious bursal disease virus (IBDV) strains were inoculated both intranasally and by eye drop into 5-week-old specific pathogen free chickens. The bursa, the liver, the kidney, the spleen, the thymus, the caecal tonsil and the thigh muscle were harvested at 4, 8, 16, 28, 40, 56, 72, 96 h post-inoculation (p.i.) for IBDV detection by in situ reverse transcriptase-polymerase chain reaction and, at the same time, the pathological changes in these tissues were investigated. A typical positive signal was detected in the liver, the kidney and the spleen of chickens inoculated with the very virulent IBDV H strain at 4 h p.i., but not in the thymus, the caecal tonsil or the thigh muscle until 8 h p.i. Virus was also found in the liver, the spleen, the kidney, the thymus, the caecal tonsils and the muscle of birds inoculated with the cell-adapted Ts strain at 4 h p.i. A positive signal was observed in the bursa later than in the other tissues. The signals increased markedly at 8 h p.i. A decrease in bursal lymphocytes was observed in haematoxylin and eosin stained sections at 28 h p.i. for the H strain and at 40 h p.i. for the Ts strain.

Animals↗
PubMed2001

Immune modulation in suckling rat pups by a growth factor extract derived from milk whey.

Oral tolerance to foreign enteral antigens is not fully developed in early neonatal life. Epidemiological evidence supports a role for maternal milk in the development of immune responses, including oral tolerance. Formula fed infants have an increased susceptibility to food allergy and the later development of autoimmune disease. This may relate to the lack in infant formula of growth factors found in maternal milk. Bovine milk contains proteins, growth factors and cytokines. Various studies have outlined the immune modulating potential of bovine milk-derived products. Fractionated whey extracts have therapeutic potential in disease states where there is an excessive inflammatory reaction, and disease preventive potential for infants who are not breast-fed. We have shown that daily oral administration of a growth factor-enriched fraction from milk whey to naturally suckling rat pups between days 4-9 postnatal can down-regulate immune activation to a specific orally administered food antigen, ovalbumin, assessed by lymphocyte proliferation. In addition, non-specific down-regulation in the intestine was observed as assessed by the expression of MHC I. Treatment of rat pups with whey extract at the time of oral sensitisation to ovalbumin also resulted in an increased secretion of TGF-beta into the culture supernatant of spleen cells incubated with specific antigen. TGF-beta is an immuno-down-regulatory cytokine involved in tolerance induction. Immune modulation by extracts derived from milk whey could be of potential benefit for formula-fed and pre-term infants in reducing susceptibility to inappropriate activation to food antigens.

Animals↗
PubMed2001

CD40 and CD86 upregulation with divergent CMRF44 expression on blood dendritic cells in inflammatory bowel diseases.

OBJECTIVE: Dendritic cells (DC) are the only antigen-presenting cells that can activate naïve T lymphocytes and initiate a primary immune response. They are also thought to have a role in immune tolerance. DC traffic from the blood to peripheral tissue where they become activated. They then present antigen and the costimulating signals necessary to initiate an immune response. In this study, we investigated the number, subsets, and activation pattern of circulating and intestinal DC from patients with clinically mild ulcerative colitis (UC) or Crohn's disease. METHODS: Patients were recruited, if they were not taking immunosuppressive therapy, and were assessed for clinical severity of their disease using for UC, the Clinical Activity Index, and for Crohn's disease, the Crohn's Disease Activity Index. Blood CD11c+ and CD11c- DC subsets, expression of costimulatory antigens, CD86 and CD40, and the early differentiation/activation antigen, CMRF44, were enumerated by multicolor flow cytometry of lineage negative (lin- = CD3-, CD19-, CD14-, CD16-) HLA-DR+ DC. These data were compared with age-matched healthy and the disease control groups of chronic noninflammatory GI diseases (cGI), acute noninflammatory GI diseases (aGI), and chronic non-GI inflammation (non-GI). In addition, cryostat sections of colonoscopic biopsies from healthy control patients and inflamed versus noninflamed gut mucosa of inflammatory bowel disease (IBD) patients were examined for CD86+ and CD40+ lin- cells. RESULTS: Twenty-one Crohn's disease and 25 UC patients, with mean Crohn's Disease Activity Index of 98 and Clinical Activity Index of 3.1, and 56 healthy controls, five cGI, five aGI, and six non-GI were studied. CD11c+ and CD11c- DC subsets did not differ significantly between Crohn's, UC, and healthy control groups. Expression of CD86 and CD40 on freshly isolated blood DC from Crohn's patients appeared higher (16.6%, 31%) and was significantly higher in UC (26.6%, 46.3%) versus healthy controls (5.5%, 25%) (p = 0.004, p = 0.012) and non-GI controls (10.2%, 22.8%) (p = 0.012, p = 0.008), but not versus cGI or aGI controls. CD86+ and CD40+ DC were also present in inflamed colonic and ileal mucosa from UC and Crohn's patients but not in noninflamed IBD mucosa or normal mucosa. Expression of the CMRF44 antigen was low on freshly isolated DC, but it was upregulated after 24-h culture on DC from all groups, although significantly less so on DC from UC versus Crohn's or healthy controls (p = 0.024). The CMRF44+ antigen was mainly associated with CD11c+ DC, and in UC was inversely related to the Clinical Activity Index (r = -0.69, p = 0.0002). CONCLUSIONS: There is upregulation of costimulatory molecules on blood DC even in very mild IBD but surprisingly, there is divergent expression of the differentiation/activation CMRF44 antigen. Upregulation of costimulatory molecules and divergent expression of CMRF44 in blood DC was also apparent in cGI and aGI but not in non-GI or healthy controls, whereas intestinal CD86+ and CD40+ DC were found only in inflamed mucosa from IBD patients. Persistent or distorted activation of blood DC or divergent regulation of costimulatory and activation antigens may have important implications for gut mucosal immunity and inflammation.

Adolescent↗
PubMed2000

Effect of lanthanum on ion absorption in cucumber seedling leaves.

Scanning electron microscope and energy-dispersive X-ray analysis were used to study the tissular distributions of elements Na, Mg, Cl, K, Ca, Mn, and Fe in leaves of cucumber seedlings in the absence or presence of La3+. The results showed that the atomic percentages of Na, Mg, Cl, K, and Ca were basically reduced and those of Mn and Fe were increased in the presence of La3+; in addition, at 0.02 mM La3+, the reduced or increased degrees were higher than those at 2.0 mM La3+. The effects of La3+ on ion absorption were similar to those of Ca2+, suggesting that the rare earth element lanthanum affects the plant physiological mechanism by regulating the Ca2+ level in plant cell.

Cucumis sativus↗
PubMed2000

[Changes of femur minerals and serum BGP in hindlimb unloaded rats during convalescence].

Objective. To observe bone mass changes during convalescence after simulated weightlessness. Method. 7-week-old rats were tail-suspended for 21 d then reloaded for 7 d and 21 d to recover, and measured serum BGP. Result. Tail suspension of rats for 21 d caused significant decrease of serum BGP and phosphorus as well as femur minerals. Serum BGP and femur minerals were still lower than control levels, but serum contents of calcium, phosphorus and magnesium increased significantly after reloading for 7 d. Femur minerals and serum BGP, calcium, phosphorus and magnesium returned to control levels after reloading for 21 d. Conclusion. The deficit in femur mineral induced by hindlimb unloading in rats can be restored by return to normal weight bearing, BGP can be used to monitor the case of its recovery.

Animals↗
PubMed1999

Sensitive and selective liquid chromatographic assay of memantine in plasma with fluorescence detection after pre-column derivatization.

A procedure was developed for the determination of memantine in plasma using liquid chromatography with fluorescence detection. Following a liquid-liquid extraction from 1 ml of plasma containing the internal standard amantadine, the extract was derivatized at room temperature with dansyl chloride, and the highly fluorescent derivatives were chromatographed with a reversed-phase C18 column and a mobile phase of phosphate buffer and acetonitrile. Dansylated memantine and amantadine were eluted in less than 13 min with no interference from endogenous material. The calibration curve was linear over the concentration range of 3-400 ng/ml with inter- and intra-assay imprecision (C.V.) of less than 10%. The limit of quantitation was 3 ng/ml, and no major antidepressant, neuroleptic or their respective metabolites interfered with the quantitation of memantine. This method could also be applied to the quantitation of amantadine.

Excitatory Amino Acid Antagonists↗
PubMed1999

The effect of lanthanide chloride on abscisic acid and electron-transport activity of some crops.

The abscisic acid (ABA) content of the root tips of four crops grown in lanthanide chloride solution and their root lengths had been determined. At lanthanide concentrations of 5 and 10 ppm, these crops all grew well and the ABA decreased. At higher lanthanide concentrations (100-500 ppm), the ABA is increased again. At these concentrations of lanthanum chloride, the photosystems I and II (PSI and PSII) and whole electron chain transport activities were inhibited. PSII was more sensitive than PSI, and it is concluded that La3+ acts on the diphenylcarbazide (DPC) action place of PSII oxidizing site.

Abscisic Acid↗
PubMed1999

Localization of transforming growth factor-beta receptor types I, II, and III in the postnatal rat small intestine.

Transforming growth factor-beta2 (TGF-beta2) levels in rat milk are high in early lactation, whereas endogenous TGF-beta1 expression in the neonatal gut increases toward midweaning. Three types of transmembrane TGF-beta receptors have been identified in mammals. The receptor III (or betaglycan) binds and presents TGF-beta1 or beta2 to receptor II. Receptor I then interacts with receptor II, forming a signaling receptor complex, and propagates the signal. To determine whether TGF-beta receptor expression in the gut is also developmentally regulated, the present study assessed ontogeny of TGF-beta receptor expression in the postnatal rat small intestine. Jejunum and ileum tissues from rat pups at d 3, 10, 14, 21, and 28 of age were collected. Cryostat sections were stained with antibodies against TGF-bea receptors I, II, and III, and various cell markers by immunofluorescence. In both regions, receptor I staining was seen on apical and basolateral membranes of the villus and crypt epithelium at all ages, and staining on the apical membrane increased with age; receptor II was predominantly expressed in the crypt, and staining on the villi appeared after d 10; receptor III was distributed throughout the mucosa at early ages but diminished from the epithelium postweaning by d 28. T cells, B cells, and dendritic cells in the lamina propria expressed TGF-beta receptor III but lacked expression of receptor I and II. The pattern of TGF-beta receptor expression changes with age in a manner that may reflect the change in ligand from TGF-beta2 (milk-derived) to TGF-beta1 (endogenously produced).

Animals↗
PubMed1998

Transforming growth factor-beta levels in maternal milk and expression in postnatal rat duodenum and ileum.

After birth, the gastrointestinal tract of the neonate is exposed to food and bacterial and environmental antigens. Maternal milk components may play a role in regulation of mucosal immune activity to luminal antigens. In this study we determine the ontogeny of transforming growth factor (TGF)-beta1-producing cells in the rat pup small intestine and assess maternal milk concentrations of TGF-beta. Intestinal tissue samples of duodenum and ileum were collected, processed, and stained for TGF-beta1, and in situ hybridization for TGF-beta1 mRNA was also performed on the duodenum. TGF-beta levels in milk were assayed by ELISA. TGF-beta2 levels in milk were high at d 6, and declined thereafter at d 10 and 19. TGF-beta1 was not detected. In contrast, the cell number and intensity of staining of TGF-beta1 peptide in the small intestine was low in 3- and 10-d-old rats and increased markedly by 19 d of life. In the duodenum mRNA levels mirrored this trend. TGF-beta1 expression in the lamina propria was absent before d 19, and increased progressively over time. Maternal milk TGF-beta2 levels are high in early milk and decrease during the weaning period. In contrast, endogenous TGF-beta production in the small intestine increases during the weaning period.

Aging↗
PubMed1993

[Pharmacological study on spleen-stomach warming and analgesic action of Cinnamomum cassia Presl].

This study deals with the antiulcer effect of water extract and ether extract of Cinnamomum cassia on four types of experimental gastric ulcer and with the antidiarrhea effect on two types of medicine-induced diarrhea in mice. These extracts have choleretic effect in anesthetized rats, and are analgesic as well. This is the pharmacologic basis of spleen-stomach warming and analgesic action of Cinnamomum cassia.

Animals↗
PubMed1992

Chemical synthesis, molecular cloning, and expression of the gene coding for the Trichosanthes trypsin inhibitor--a squash family inhibitor.

The gene coding for a Trichosanthes trypsin inhibitor analog (Ala-6-TTI) in which methionine at position 6 was replaced by alanine was synthesized chemically. The synthetic gene was cloned into plasmid pWR590-1 and expressed in Escherichia coli as a fusion protein composed of beta-galactosidase fragment of 590 amino acid residues and (Ala-6)-TTI, with methionine as a connecting residue. After cyanogen bromide cleavage and reduction of the fusion protein, followed by refolding with trypsin-Sepharose 4B as a matrix and affinity chromatography on the immobilized enzyme, the fully active (Ala-6)-TTI was obtained. The trypsin inhibitory activity and amino acid composition of the recombinant (Ala-6)-TTI were consistent with those of the natural one. The (Ala-6)-TTI gene was also cloned into the secretion expression vector, pVT102U/alpha, in Saccharomyces cerevisiae. In order to make the reading frame of the gene compatible with the vector, a nucleotide was inserted into the (Ala-6)-TTI gene via site-directed mutagenesis. The secreted (Ala-6)-TTI was purified and found to be correctly processed at the junction between the alpha-factor leader peptide and (Ala-6)-TTI downstream. Of the two expression systems, the latter is more advantageous in the high yield (greater than 2 mg/liter), easy purification and needlessness of disulfide refolding.

Amino Acid Sequence↗
PubMed1992

Sensitive and selective liquid-chromatographic assay of fluoxetine and norfluoxetine in plasma with fluorescence detection after precolumn derivatization.

We determined fluoxetine (Prozac) and its major metabolite norfluoxetine in plasma by liquid chromatography with fluorescence detection. After liquid-liquid extraction from 1 mL of plasma, the extract was derivatized at room temperature with dansyl chloride, and the highly fluorescent derivatives were chromatographed with a reversed-phase C18 column and a mobile phase of phosphate buffer and acetonitrile. Dansylated fluoxetine, norfluoxetine, and the internal standard were eluted in less than 14 min with no interference from endogenous material. The calibration curve was linear over the concentration range 25-800 micrograms/L with inter- and intra-assay imprecision (CV) of less than 10%. Validity of the assay was checked by comparing results for 110 patients' samples with those by a liquid-chromatographic method with ultraviolet detection (r = 0.993 for fluoxetine, 0.957 for norfluoxetine). The identity of the dansylated derivatives was verified by positive chemical ionization mass spectroscopy. The lower limit of detection was approximately 3 micrograms/L. Because no major antidepressant, neuroleptic, or respective drug metabolites interfere with the quantification of fluoxetine and norfluoxetine, this is a useful procedure for pharmacokinetic studies and in clinical settings.

Chromatography, Liquid↗